Team:KIT-Kyoto/kazukokekokko
From 2012.igem.org
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BP reaction by invitrogen gateway system1. PCR using promers containing the attB sequence. *attB sequence: 2. Purification PCR products. 3. Prepare vials in 1.5mL tube according to the following components.
4. Dissolve BP Clonase Ⅱ enzyme mix on ice. 5. Add BP Clonase Ⅱ enzyme mix (2uL or 1uL) to vials, vortex and spin them down. 6. Incubate vials at 25℃ for more than an hour. 7. Add Proteinase K solutionto vials (1uL) and vortex shortly. 8. Incubate at 37℃ for 10 minutes. LR reaction by invitrogen gateway system1. Control vials in 1.5mL tube according to the following components.
2. Dissolve LR Clonase Ⅱ enzyme mix on ice 3. Add LR Clonase Ⅱ enzyme mix to vials by 2uL or 1uL, votex and spin down. 4. Incubate vials at 25℃ for more than an hour. 5. Add Proteinase K solution to vials by 1uL and vortex it shortly. 6. Incubate at 37℃ for 30 minutes. |
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